Fig. 2.

MAP3K1 and MAP2K4 knockout confer sensitivity to selumetinib. a MAP3K1 or MAP2K4 knockout cells were generated using a lentiviral CRISPR/Cas9 vector. Control and MAP3K1 or MAP2K4 knockout MDA-MB-468, H358 or HCT116 cells were treated with 0.02 μM trametinib for 72 h, and lysates were western blotted for MAP3K1 and MAP2K4. HSP90 served as a control. b, c Control and MAP3K1 or MAP2K4 knockout MDA-MB-468, H358 or HCT116 cells were cultured for 2 weeks in medium containing the indicated concentration of selumetinib. Then cells were fixed and stained. d Control and MAP2K4 knockout LoVo or DLD1 cells were treated with 2 μM selumetinib, lysates were western blotted for MAP2K4. HSP90 served as a control. e Control and MAP2K4 knockout LoVo or DLD1 cells were cultured for two weeks in medium containing the indicated concentration of selumetinib. Then cells were fixed and stained. f, g Control and MAP3K1 or MAP2K4 knockout MDA-MB-468, H358 or HCT116 cells were cultured for two weeks in medium containing the indicated concentration of trametinib (f) or SCH772984 (g). Then cells were fixed and stained